The cell invasion and migration assays were performed using Transwell chambers (Corning, Inc.), with or without Matrigel (BD Biosciences), respectively. Briefly, 5×104 or 1×105 transfected cells were suspended in serum-free medium and plated in the upper chamber for the migration and invasion assays, respectively. The medium in the lower chamber was supplemented with 10% FBS. After 16 or 24 h, cells in the lower chamber were fixed in 10% formalin at 25°C for 10 min and stained with 0.1% crystal violet at 25°C for 5 min and counted using an inverted light microscope at ×100 magnification (Olympus Corporation).

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